Creating hybridomas - A Guide to Generating Antibody-Producing Cell Lines摘要:Creating hybridomas - A Guide to Generating Antibody-Producing Cell LinesIntroduction
Creating hybridoma cells is an indispensable technique widely used in the
Introduction
Creating hybridoma cells is an indispensable technique widely used in the field of immunology to produce monoclonal antibodies with high specificity and affinity. This technique is based on the fusion of myeloma cells and antibody-secreting plasma cells, resulting in the formation of hybridomas. In this article, we will explore the steps involved in generating hybridomas through the fusion of immune cells with myeloma cells.Generation of Immune Cells and Myeloma Cells
The very first step in generating hybridomas is the preparation of immune cells and myeloma cells. Immune cells are obtained from animals by triggering an immune response against the antigen of interest. After obtaining the splenocytes or lymphocytes, the immune cells are typically purified by density gradient centrifugation. In contrast, myeloma cells, which are taken from immortalized myeloma cell lines, are cultured separately.Hybridoma Formation
The next step is the fusion of immune cells and myeloma cells, which is typically achieved using polyethylene glycol (PEG). The primary goal of this step is to initiate the fusion process by causing the plasma cells and myeloma cells to merge. After the fusion, the hybridomas are recovered using a selective medium containing hypoxanthine, aminopterin, and thymidine (HAT medium). In this culture medium, only the fused hybridoma cells can survive, as the growth of plasma cells and myeloma cells is inhibited due to their lack of key enzymes for nucleotide synthesis.Screening of Hybridomas
After the generation of hybridoma cells, the next step is to screen the cells to identify clones that produce antibodies targeting the antigen of interest. ELISA or flow cytometry can be used to screen hybridoma cells. Once identified, the antibodies produced by the hybridomas can be purified from the cell culture supernatant using various chromatographic techniques.Conclusion
In conclusion, the generation of hybridomas is a crucial technique for producing monoclonal antibodies with high specificity and affinity in research and diagnostic applications. These steps, although challenging, can be accomplished with the desired outcome by following the optimized protocol for generating hybridoma cells. The potential availability of these antibody-producing hybridomas in unlimited quantities makes them a valuable resource in the field of biomedical research.版权声明:本站部分常识内容收集于其他平台,若您有更好的常识内容想分享可以联系我们哦!